Department of Endocrinology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Research Centre for Clinical Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Front Endocrinol (Lausanne). 2021 Nov 18;12:731107. doi: 10.3389/fendo.2021.731107. eCollection 2021.
Androgen insensitivity syndrome (AIS) is a rare X-linked genetic disease and one of the causes of 46,XY disorder of sexual development. The unstraightforward diagnosis of AIS and the gender assignment dilemma still make a plague for this disorder due to the overlapping clinical phenotypes.
Peripheral blood mononuclear cells (PBMCs) of partial AIS (PAIS) patients and healthy controls were separated, and RNA-seq was performed to investigate transcriptome variance. Then, tissue-specific gene expression, functional enrichment, and protein-protein interaction (PPI) network analyses were performed; and the key modules were identified. Finally, the RNA expression of differentially expressed genes (DEGs) of interest was validated by quantitative real-time PCR (qRT-PCR).
In our dataset, a total of 725 DEGs were captured, with functionally enriched reproduction and immune-related pathways and Gene Ontology (GO) functions. The most highly specific systems centered on hematologic/immune and reproductive/endocrine systems. We finally filtered out CCR1, PPBP, PF4, CLU, KMT2D, GP6, and SPARC by the key gene clusters of the PPI network and manual screening of tissue-specific gene expression. These genes provide novel insight into the pathogenesis of AIS in the immune system or metabolism and bring forward possible molecular markers for clinical screening. The qRT-PCR results showed a consistent trend in the expression levels of related genes between PAIS patients and healthy controls.
The present study sheds light on the molecular mechanisms underlying the pathogenesis and progression of AIS, providing potential targets for diagnosis and future investigation.
雄激素不敏感综合征（AIS）是一种罕见的 X 连锁遗传性疾病，也是 46,XY 性发育障碍的原因之一。由于重叠的临床表型，AIS 的诊断不明确以及性别分配困境仍然是该疾病的难题。
分离部分 AIS（PAIS）患者和健康对照者的外周血单核细胞（PBMCs），进行 RNA-seq 以研究转录组差异。然后进行组织特异性基因表达、功能富集和蛋白质-蛋白质相互作用（PPI）网络分析，并鉴定关键模块。最后，通过定量实时 PCR（qRT-PCR）验证感兴趣的差异表达基因（DEG）的 RNA 表达。
在我们的数据集，总共捕获了 725 个 DEG，具有丰富的功能再生和免疫相关途径和基因本体论（GO）功能。最高度特异性的系统集中在血液/免疫和生殖/内分泌系统。我们最终通过 PPI 网络的关键基因簇和组织特异性基因表达的手动筛选，筛选出 CCR1、PPBP、PF4、CLU、KMT2D、GP6 和 SPARC。这些基因提供了免疫系统或代谢中 AIS 发病机制的新见解，并提出了可能的临床筛选分子标志物。qRT-PCR 结果显示 PAIS 患者和健康对照者之间相关基因表达水平的一致趋势。
本研究阐明了 AIS 发病机制和进展的分子机制，为诊断和未来研究提供了潜在的靶点。
